This invention relates to an improved process for producing heparinase utilizing a defined medium with controlled concentrations of carbon source and sulfur source. Little or no heparin or other inducer is required in the medium for the synthesis of heparinase.
Heparinase is an enzyme presently used in assays for heparin. Presently heparinase is produced by Flavobacterium heparnium in a defined culture medium typically containing glucose, ammonium sulfate and a mixture of potassium monobasic phosphate and sodium dibasic phosphate, magnesium sulfate, trace salts, L-methionine and L-histidine and the heparinase inducer, such as sodium heparin or other salt, heparin monosulfate, hyaluronic acid, maltose, N-acetyl-D-glucosamine or the like. Certain mutants of Flavobacterium heparinum may not require a heparinase inducer in this medium.
Due to the high cost of potent heparinase inducers, such as heparin, it would be desirable as a first step to produce heparinase in this way by alteration of the bacterial growth medium rather than having to obtain mutants which produce heparinase without the inducer. Alternatively, it would be desirable to improve heparinase production by altering the growth medium in such a way as to improve the potency of the inducer and/or to improve the cell density levels achieved in the growth medium.